Western blot abreview for Anti-HA tag antibody [4C12]

Average
Abreviews
Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing (10)
Sample
Human Cell lysate - whole cell (HEK293)
Specification
HEK293
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: 5% milk
Dilution
1/1000

Secondary antibody

Name
Non-Abcam antibody was used: Jackson anti-mouse heavy and light
Host species: Goat
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase
Dilution
1/20000

Detection

Detection method
SuperSignal West Pico
Bands
Specific: 65 kDa Non-specific: 80, 90, 50 kDa
Exposure
30 second(s)
Negative control
Lane 1: Untransfected HEK293 cells.
Positive control
Lane 2: HEK293 Cells transfected with an HA- tagged protein that runs at approximately 65kDa

Additional data

Additional Notes
Strong non-specific bands in this application between 75 and 100kDa would prevent use of this antibody for HA-tagged proteins in that size range.

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Submitted Mar 03 2015

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